Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol (39.85% ±11.41) than in 5% glycerol treatment (18.08%±1.61). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol (34.12%±11.02) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into LN2. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol (72.5±5.00%, 54.88±0.66% and 46.00±2.40%; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol (34.69±4.64% vs 46.00±2.40%; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: 55.81±2.94, 55.19±3.34 vs 47.94±3.48%; p<0.05 and membrane integrity: 44.94±3.51, 46.06±2.25 vs 40.38±1.03%; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

We investigated the effects of cadmium exposure and various stress on the transcription of heat shock protein 70 and 82 (HSP70 and HSP82) from Pardosa astrigera wolf spider. To do this, P. astrigera HSP70 and HSP82 genes were cloned and its full-length sequence determined. Female spiders were long-term exposed to cadmium or to polychlorinated biphenyl (PCB) for 2, 4 and 6 weeks and short-term exposed to endosulfan by dietary uptake. Female spiders were also exposed to various temperatures. HSP82 did not show a clear tendency of transcription induction following exposure to cadmium. On the contrary, HSP70 transcription gradually increased during the exposure to 2, 20 and 40 mM of cadmium for 2, 4 and 6 weeks. Transcript level of HSP70 was not significantly changed by endosulfan and PCB exposure. In the short-term (3 hr) temperature exposure, an increased expression of HSP70 was observed under the heat shock to 30°C and then slightly decreased at 35°C. However, induction of HSP70 transcription was not observed during the long-term (7 days) temperature exposure. Taken together, HSP70 gene appears to be up-regulated by cadmium in a time-dependent manner but little affected by other potential contaminants. Analysis of HSP70 transcript levels in P. astrigera collected from various fields revealed that levels of cadmium concentration were well correlated with HSP70 transcript levels (r2 = 0.76). Taken together, it was suggested that transcript level of HSP70 could be useful as a biomarker for the long-term cadmium exposure of P. astrigera.

We examined the effects of cadmium exposure and various temperature stress on the expression of Pardosa astrigera heat shock protein 70 (HSP70). To do this, P. astrigera HSP70 gene was cloned and its sequence determined. Female spiders collected from non-contaminated region were exposed to 40mM CdCl2 for 2, 4 and 6 weeks by dietary uptake. At the end of every 2, 4 and 6 weeks of exposure, a batch of 5 spiders was collected and total RNA was extracted from each batch of whole bodies. Female spiders were also exposed to different temperatures (20, 25, 30 and 35℃) for 3h and RNA extracted likewise. Transcription profiles of HSP70 in response to cadmium and temperature were determined by quantitative real-time PCR using 18S rRNA as reference gene for data normalization. HSP70 transcription gradually increased during 2,4 and 6 weeks of exposure to cadmium. In particular, the expression level at 6-week exposure was 3.4-fold higher than that of untreated control. In the temperature response, an increased expression of HSP70 was also observed as temperature increased up to 30℃ and then slightly decreased at 35℃. The expression level at 30℃ was 2.3-fold higher than that of 25℃. Taken together, HSP70 gene appears to be up-regulated by general stress factors including cadmium exposure and temperature increase.

In this study, to investigate the effects of herb aroma components, the BDI test was performed with the 124 students of the Youngdong University and the subjects whose score was 16 or higher were selected and allocated to the herb-extracted aroma-treated group and the non-treated group, 27 and 10 students for each group. The BDI and SDS tests were carried out at each stage (before and after the treatment, 10 days later and 30 days later). The result showed that the pre-treatment BDI test result was significantly different from all those of the post-treatment test, and the tests after 10 days and 30 days in the aroma-treated group. In the SDS test, the pre-treatment test result was significantly different from the results of the post-treatment test and the test after 10 days, while it was not significantly different from the result of the test after 30 days. Additionally, to verify whether the change within the group is larger than that by natural recovery or not, ANCOVA was performed with respect to the difference in the pre-treatment test score between the groups depending on whether the treatment was given or not, having the SDS pre-treatment score as the covariate, and the result showed that the post-treatment test scores were significantly different. Thus, it was verified that, if the SDS test score is considered as the depression indicator, the effect of aromatherapy was greater than the change by natural recovery. The difference in the post-treatment test score was analyzed depending on whether the treatment was given or not, having the BDI pre-treatment test score as the control variable, and the result showed that the post-treatment test scores were not significantly different. Based on such a theoretical verification, it is assumed that the nature-friendly treatment method using herb aroma components can be a great help in suppressing depression. Therefore, it is expected that herb aroma components can provide systematic therapeutic effect on the suppression of depression.